Sex Dependent Disparities in the Central Innate Immune Response after Moderate Spinal Cord Contusion in Rat.

Subacute spinal cord injury (SCI) displays a complex pathophysiology associated with pro-inflammation and ensuing tissue damage. Microglia, the resident innate immune cells of the CNS, in concert with infiltrating macrophages, are the primary contributors to SCI-induced inflammation. However, subpopulations of activated microglia can also possess immunomodulatory activities that are essential for tissue remodeling and repair, including the production of anti-inflammatory cytokines and growth factors that are vital for SCI recovery. Recently, reports have provided convincing evidence that sex-dependent differences exist in how microglia function during CNS pathologies and the extent to which these cells contribute to neurorepair and endogenous recovery. Herein we employed flow cytometry and immunohistochemical methods to characterize the phenotype and population dynamics of activated innate immune cells within the injured spinal cord of age-matched male and female rats within the first week (7 days) following thoracic SCI contusion. This assessment included the analysis of pro- and anti-inflammatory markers, as well as the expression of critical immunomodulatory kinases, including P38 MAPK, and transcription factors, such as NFκB, which play pivotal roles in injury-induced inflammation. We demonstrate that activated microglia from the injured spinal cord of female rats exhibited a significantly diminutive pro-inflammatory response, but enhanced anti-inflammatory activity compared to males. These changes included lower levels of iNOS and TLR4 expression but increased levels of ARG-1 and CD68 in females after SCI. The altered expression of these markers is indicative of a disparate secretome between the microglia of males and females after SCI and that the female microglia possesses higher phagocytic capabilities (increased CD68). The examination of immunoregulatory kinases and transcription factors revealed that female microglia had higher levels of phosphorylated P38Thr180/Tyr182 MAPK and nuclear NFκB pp50Ser337 but lower amounts of nuclear NFκB pp65Ser536, suggestive of an attenuated pro-inflammatory phenotype in females compared to males after SCI. Collectively, this work provides novel insight into some of the sex disparities that exist in the innate immune response after SCI and indicates that sex is an important variable when designing and testing new therapeutic interventions or interpretating positive or negative responses to an intervention.

Scaling artificial light at night and disease vector interactions into socio-ecological systems: a conceptual appraisal.

There is burgeoning interest in how artificial light at night (ALAN) interacts with disease vectors, particularly mosquitoes. ALAN can alter mosquito behaviour and biting propensity, and so must alter disease transfer rates. However, most studies to date have been laboratory-based, and it remains unclear how ALAN modulates disease vector risk. Here, we identify five priorities to assess how artificial light can influence disease vectors in socio-ecological systems. These are to (i) clarify the mechanistic role of artificial light on mosquitoes, (ii) determine how ALAN interacts with other drivers of global change to influence vector disease dynamics across species, (iii) determine how ALAN interacts with other vector suppression strategies, (iv) measure and quantify the impact of ALAN at scales relevant for vectors, and (v) overcome the political and social barriers in implementing it as a novel vector suppression strategy. These priorities must be addressed to evaluate the costs and benefits of employing appropriate ALAN regimes in complex socio-ecological systems if it is to reduce disease burdens, especially in the developing world. This article is part of the theme issue 'Light pollution in complex ecological systems'.

Host brood traits, independent of adult behaviours, reduce Varroa destructor mite reproduction in resistant honeybee populations.

The ectoparasitic mite Varroa destructor is an invasive species of Western honey bees (Apis mellifera) and the largest pathogenic threat to their health world-wide. Its successful invasion and expansion is related to its ability to exploit the worker brood for reproduction, which results in an exponential population growth rate in the new host. With invasion of the mite, wild honeybee populations have been nearly eradicated from Europe and North America, and the survival of managed honeybee populations relies on mite population control treatments. However, there are a few documented honeybee populations surviving extended periods without control treatments due to adapted host traits that directly impact Varroa mite fitness. The aim of this study was to investigate if Varroa mite reproductive success was affected by traits of adult bee behaviours or by traits of the worker brood, in three mite-resistant honey bee populations from Sweden, France and Norway. The mite's reproductive success was measured and compared in broods that were either exposed to, or excluded from, adult bee access. Mite-resistant bee populations were also compared with a local mite-susceptible population, as a control group. Our results show that mite reproductive success rates and mite fecundity in the three mite-resistant populations were significantly different from the control population, with the French and Swedish populations having significantly lower reproductive rates than the Norwegian population. When comparing mite reproduction in exposed or excluded brood treatments, no differences were observed, regardless of population. This result clearly demonstrates that Varroa mite reproductive success can be suppressed by traits of the brood, independent of adult worker bees.

Reproductive success of the parasitic mite (Varroa destructor) is lower in honeybee colonies that target infested cells with recapping.

Cell recapping is a behavioural trait of honeybees (Apis mellifera) where cells with developing pupae are uncapped, inspected, and then recapped, without removing the pupae. The ectoparasitic mite Varroa destructor, unarguably the most destructive pest in apiculture world-wide, invades the cells of developing pupae to feed and reproduce. Honeybees that target mite infested cells with this behaviour may disrupt the reproductive cycle of the mite. Hence, cell recapping has been associated with colony-level declines in mite reproduction. In this study we compared the colony-level efficacy of cell recapping (how often infested cells are recapped) to the average mite fecundity in A. mellifera. Our study populations, known to be adapted to V. destructor, were from Avignon, France, Gotland, Sweden, and Oslo, Norway, and were compared to geographically similar, treated control colonies. The results show that colonies with a higher recapping efficacy also have a lower average mite reproductive success. This pattern was likely driven by the adapted populations as they had the largest proportion of highly-targeted cell recapping. The consistent presence of this trait in mite-resistant and mite-susceptible colonies with varying degrees of expression may make it a good proxy trait for selective breeding on a large scale.

Author Correction: Structural basis of indisulam-mediated RBM39 recruitment to DCAF15 E3 ligase complex.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Structural basis of indisulam-mediated RBM39 recruitment to DCAF15 E3 ligase complex.

The anticancer agent indisulam inhibits cell proliferation by causing degradation of RBM39, an essential mRNA splicing factor. Indisulam promotes an interaction between RBM39 and the DCAF15 E3 ligase substrate receptor, leading to RBM39 ubiquitination and proteasome-mediated degradation. To delineate the precise mechanism by which indisulam mediates the DCAF15-RBM39 interaction, we solved the DCAF15-DDB1-DDA1-indisulam-RBM39(RRM2) complex structure to a resolution of 2.3 Å. DCAF15 has a distinct topology that embraces the RBM39(RRM2) domain largely via non-polar interactions, and indisulam binds between DCAF15 and RBM39(RRM2), coordinating additional interactions between the two proteins. Studies with RBM39 point mutants and indisulam analogs validated the structural model and defined the RBM39 α-helical degron motif. The degron is found only in RBM23 and RBM39, and only these proteins were detectably downregulated in indisulam-treated HCT116 cells. This work further explains how indisulam induces RBM39 degradation and defines the challenge of harnessing DCAF15 to degrade additional targets.

Metabolic rate does not vary with seasonal change in Anopheles arabiensis adults in South Africa.

An important component of South Africa's malaria elimination agenda is identifying the entomological drivers of residual transmission, especially those that present opportunities for enhanced vector control. Seasonal mosquito density correlates directly with malaria transmission in South Africa. Transmission is highest during the warm rainy season and lowest but not entirely absent during the cooler dry season. The factors that sustain dry-season mosquito survival remain unknown. The aim of this project was therefore to investigate seasonal change in metabolic rate to determine the presence or absence of winter dormancy in malaria vector mosquitoes. Metabolic rate, determined by CO2 production during closed-system respirometry, was measured from wild anophelines collected from KwaZulu-Natal Province, South Africa. Monthly sampling spanned all four seasons (summer, autumn, winter, and spring) in 2017. Anopheles arabiensis and An. parensis specimens formed the majority of the total 437 identified specimens (n = 216 and n = 162, respectively). Metabolic rate data from wild-caught mosquitoes showed no significant seasonal disparities for An. arabiensis and An. parensis males and females. Further laboratory experiments assessed the effect of manipulated photoperiod, representing seasonal day-length changes, on the metabolic rate of colonized An. arabiensis mosquitoes. Simulations of midwinter (10 h:14 h light dark) and midsummer (14 h:10 h) day-length showed no significant effect on the metabolic rate of these mosquitoes. Age (in days) had a significant effect on the metabolic rate of both male and female colonized adult An. arabiensis mosquitoes which may be linked to developmental factors during maturation of adults. These data suggest that the South African populations of the malaria vector species An. arabiensis and An. parensis do not curtail their breeding and foraging activities during the colder and drier winter months. Overwintering by diapause does not appear to be triggered in the adult mosquito stage in An. arabiensis. However, their respective population densities do decrease considerably during winter leading to reduced malaria transmission and the opportunity for control by winter larviciding of known breeding sites.

Anopheles parensis contributes to residual malaria transmission in South Africa.

BACKGROUND: Understanding the contribution of outdoor-resting Anopheles mosquitoes to residual malaria transmission is important in terms of scaling up vector control towards malaria elimination in South Africa. The aim of this project was to assess the potential role of Anopheles parensis and other Anopheles species in residual malaria transmission, using sentinel surveillance sites in the uMkhanyakude District of northern KwaZulu-Natal Province. METHODS: Monthly vector surveillance was conducted at the sentinel sites from January 2017 to May 2018. Outdoor-placed clay pot resting traps were used to collect male and female adult Anopheles mosquitoes. All Anopheles gambiae complex and Anopheles funestus group specimens collected were identified to species and all females were screened for Plasmodium falciparum circumsporozoite protein (CSP) by enzyme-linked immunosorbent assay (ELISA). Samples showing infectivity for P. falciparum were further verified by a nested PCR and subsequent DNA sequence analysis. RESULTS: From a sample of 491 anophelines, Anopheles arabiensis (n = 228) and An. parensis (n = 194) were the most abundant. Other species collected included Anopheles merus (n =11), Anopheles quadriannulatus (n = 10), Anopheles leesoni (n = 29), Anopheles rivulorum (n =18), and Anopheles vaneedeni (n =1). Of the 317 female specimens screened for P. falciparum CSP, one Anopheles arabiensis and one An. parensis showed positive by ELISA and Plasmodium nested PCR. For the An. parensis specimen, confirmation of its species identity was based on sequence analysis of the ITS2 region, and the presence of P. falciparum DNA was further confirmed by sequence analysis. CONCLUSIONS: Anopheles parensis is a potential vector of malaria in South Africa although its contribution to transmission is likely to be minimal at best owing to its strong zoophilic tendency. By contrast, An. arabiensis is a major vector that is primarily responsible for the bulk of residual malaria transmission in South Africa. As all recently collected sporozoite-positive Anopheles mosquitoes were found in outdoor-placed resting traps, it is necessary to introduce interventions that can be used to control outdoor-resting vector populations while maintaining the efficacy of South Africa's indoor house spraying operations.

Aged blood impairs hippocampal neural precursor activity and activates microglia via brain endothelial cell VCAM1.

An aged circulatory environment can activate microglia, reduce neural precursor cell activity and impair cognition in mice. We hypothesized that brain endothelial cells (BECs) mediate at least some of these effects. We observe that BECs in the aged mouse hippocampus express an inflammatory transcriptional profile with focal upregulation of vascular cell adhesion molecule 1 (VCAM1), a protein that facilitates vascular-immune cell interactions. Concomitantly, levels of the shed, soluble form of VCAM1 are prominently increased in the plasma of aged humans and mice, and their plasma is sufficient to increase VCAM1 expression in cultured BECs and the hippocampi of young mice. Systemic administration of anti-VCAM1 antibody or genetic ablation of Vcam1 in BECs counteracts the detrimental effects of plasma from aged individuals on young brains and reverses aging aspects, including microglial reactivity and cognitive deficits, in the brains of aged mice. Together, these findings establish brain endothelial VCAM1 at the blood-brain barrier as a possible target to treat age-related neurodegeneration.

Bifunctional Molecular Probes for Activity-Based Visualization of Quinone-Dependent Amine Oxidases.

The design, synthesis, and evaluation of two bifunctional molecular probes that can be used to visualize quinone-dependent amine oxidase enzymes in an activity-dependent manner are described. These probes use alkylhydrazines to irreversibly bind the target enzymes, which can then be visualized with either Western blotting or in-gel fluorescence. The results show that the Western blotting readout, which utilizes commercially available anti-nitrophenyl antibodies to detect a simple dinitrophenyl antigen, provides a stronger readout than the fluorescein-based fluorescence readout. This visualization strategy can be used to measure the potency of enzyme inhibitors by selectively visualizing the active enzyme that remains after treatment with an inhibitor. Looking forward, this probe molecule and visualization strategy will enable activity-based protein-profiling experiments, such as determining inhibitor selectivity values within full proteome mixtures, for this family of amine oxidase enzymes.

A new malaria vector mosquito in South Africa.

South Africa aims to eliminate malaria within its borders by 2018. Despite well-coordinated provincial vector control programmes that are based on indoor residual insecticide spraying, low-level residual malaria transmission continues in the low-altitude border regions of the north-eastern sector of the country. In order to identify the underlying causes of residual transmission, an enhanced vector surveillance system has been implemented at selected sites in the Mpumalanga and KwaZulu-Natal (KZN) provinces. The collection periods for the data presented are March 2015 to April 2016 for Mpumalanga and January 2014 to December 2015 for KZN. The mosquito collection methods used included indoor and outdoor traps based on the use of traditional ceramic pots, modified plastic buckets and exit window traps (KZN only). All Anopheles funestus species group mosquitoes collected were identified to species and all females were screened for the presence of Plasmodium falciparum sporozoites. Two An. vaneedeni females, one from each surveillance site, tested positive for P. falciparum sporozoites. These are the first records of natural populations of An. vaneedeni being infective with P. falciparum. As both specimens were collected from outdoor-placed ceramic pots, these data show that An. vaneedeni likely contributes to residual malaria transmission in South Africa.

Comparing hydrazine-derived reactive groups as inhibitors of quinone-dependent amine oxidases.

Lysyl oxidase has emerged as an important enzyme in cancer metastasis. Its activity has been reported to become upregulated in several types of cancer, and blocking its activity has been shown to limit the metastatic potential of various cancers. The small-molecules phenylhydrazine and ß-aminopropionitrile are known to inhibit lysyl oxidase; however, issues of stability, toxicity, and poorly defined mechanisms limit their potential use in medical applications. The experiments presented herein evaluate three other families of hydrazine-derived compounds - hydrazides, alkyl hydrazines, and semicarbazides - as irreversible inhibitors of lysyl oxidase including determining the kinetic parameters and comparing the inhibition selectivities for lysyl oxidase against the topaquinone-containing diamine oxidase from lentil seedlings. The results suggest that the hydrazide group may be a useful core functionality that can be developed into potent and selective inhibitors of lysyl oxidase and eventually find application in cancer metastasis research.

Two in one: cryptic species discovered in biological control agent populations using molecular data and crossbreeding experiments.

There are many examples of cryptic species that have been identified through DNA-barcoding or other genetic techniques. There are, however, very few confirmations of cryptic species being reproductively isolated. This study presents one of the few cases of cryptic species that has been confirmed to be reproductively isolated and therefore true species according to the biological species concept. The cryptic species are of special interest because they were discovered within biological control agent populations. Two geographically isolated populations of Eccritotarsus catarinensis (Carvalho) [Hemiptera: Miridae], a biological control agent for the invasive aquatic macrophyte, water hyacinth, Eichhornia crassipes (Mart.) Solms [Pontederiaceae], in South Africa, were sampled from the native range of the species in South America. Morphological characteristics indicated that both populations were the same species according to the current taxonomy, but subsequent DNA analysis and breeding experiments revealed that the two populations are reproductively isolated. Crossbreeding experiments resulted in very few hybrid offspring when individuals were forced to interbreed with individuals of the other population, and no hybrid offspring were recorded when a choice of mate from either population was offered. The data indicate that the two populations are cryptic species that are reproductively incompatible. Subtle but reliable diagnostic characteristics were then identified to distinguish between the two species which would have been considered intraspecific variation without the data from the genetics and interbreeding experiments. These findings suggest that all consignments of biological control agents from allopatric populations should be screened for cryptic species using genetic techniques and that the importation of multiple consignments of the same species for biological control should be conducted with caution.